Abstract
A recombinant single chain fragment variable (scFv) antibody was constructed from hybridoma cell lines expressing anti-solamargine (As) monoclonal antibody (MAb). The characteristics of the As-scFv protein, expressed in both recombinant Escherichia coli and hairy root, were almost same with those of the original MAb. Up to 220 ng recombinant As-scFv was produced per milligram of soluble protein in transgenic hairy root cultures of S. khasianum. The solasodine glycosides concentration was 2.3 fold higher in the transgenic, than in the wild-type hairy root, reflecting from the soluble As-scFv level. The recombinant S. khasianum plant regenerated from recombinant hairy roots contained 2.3 fold concentration of solasodine glycoside, compared to the regenerated plant from wild-type hairy root. These results suggested that the scFv antibody expressed in transgenic plant controlled the antigen level. This methodology is the first success for molecular breeding of secondary metabolites, without understanding of biosynthetic enzyme.
Highlights
In our former stage of medicinal plant breeding, we succeeded to clonally propagate Pinellia ternate (Araceae) via protocorm-like body [1], and by liquid phase culturing, which theoretically produced 4×1023 clonal plants from a single tuber in a year
We postulate that the recombinant single chain fragment variable (scFv) can target small antigens like solasodine glycosides, which can lead to the accumulation of antigen-antibody complexes in plant organs
The molecular weight of As-scFv protein produced by transgenic hairy roots was the same as that of the protein expressed in E. coli, having 31 kD as indicated in Figure 3, whereas wild type hairy roots did not produce As-scFv [25]
Summary
In our former stage of medicinal plant breeding, we succeeded to clonally propagate Pinellia ternate (Araceae) via protocorm-like body [1], and by liquid phase culturing, which theoretically produced 4×1023 clonal plants from a single tuber in a year. We confirmed the relation between the virus infection and the component concentration, comparing with the virus free plant obtained by a tip tissue culture of Rhemannia species [6] (Scrophulariaceae). In the second stage of medicinal plant breeding, we did make evident that the medicinal plants clonally propagated promoted the homogeneity of bioactive component concentration, compared to the parent plant for aconitinetype alkaloid [7] (Aconitum spp.), atractylon [8] (Atractylodes spp.), and gentiopicroside [9] (Gentiana spp.). Solanum species belonging to Solanaceae family contain steroidal alkaloid glycoside like solasonine, solamargine and khasianine (Figure 1)
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